Reprogramming avocado lipid biosynthesis in moss for sustainable biomass oil production
Abstract
Triacylglycerols (TAGs), the predominant storage lipids in plants, are central to vegetable oil production. With global demand for plant-derived oils projected to increase by 2030, there is a need for sustainable platforms that can produce oils beyond seed-based agriculture. This study aims to metabolically engineer moss (Physcomitrium patens) as a genetically tractable chassis for enhanced lipid production using avocado-derived transcription factors and enzymes. We successfully demonstrated a significant increase in TAG content, through transient co-expression of four avocado (Persea americana) genes, namely WRINKLED1 (WRI1), WRI2, diacylglycerol acyltransferases 1 (DGAT1), and phospholipid: diacylglycerol acyltransferase 1 (PDAT1) in Nicotiana benthamiana leaves. We hypothesize that stable expression of these “push–pull” avocado module genes in moss will increase storage lipid synthesis and redirect carbon flux toward TAG in vegetative biomass. Avocado cDNAs encoding the transcription factors PaWRI1 and PaWRI2 were cloned into Gateway-compatible expression vectors and introduced into moss protoplasts via PEG-mediated transformation. Stable transgenic moss lines expressing PaWRI1 and PaWRI2 were successfully generated. Total lipids from wild-type gametophores were extracted and analyzed by thin-layer chromatography (TLC) and gas chromatography–mass spectrometry (GC–MS) to establish a baseline lipid profile. Ongoing work is focused on evaluating TAG accumulation in the transgenic lines, as well as visualizing and quantifying lipid droplet formation using Nile Red staining and confocal fluorescence microscopy. In parallel, the avocado acyltransferase genes PaDGAT1 and PaPDAT1 are being cloned into Gateway-compatible expression vectors for subsequent introduction into moss, enabling future stacking of the full four-gene avocado lipid-biosynthesis module to further enhance TAG production. Collectively, these approaches will establish moss as a controllable, vegetative platform for plant oil engineering.
Start Time
15-4-2026 9:00 AM
End Time
15-4-2026 10:00 AM
Room Number
311
Presentation Type
Oral Presentation
Presentation Subtype
Grad/Comp Orals
Presentation Category
Science, Technology, and Engineering
Student Type
Graduate
Faculty Mentor
Aruna Kilaru
Reprogramming avocado lipid biosynthesis in moss for sustainable biomass oil production
311
Triacylglycerols (TAGs), the predominant storage lipids in plants, are central to vegetable oil production. With global demand for plant-derived oils projected to increase by 2030, there is a need for sustainable platforms that can produce oils beyond seed-based agriculture. This study aims to metabolically engineer moss (Physcomitrium patens) as a genetically tractable chassis for enhanced lipid production using avocado-derived transcription factors and enzymes. We successfully demonstrated a significant increase in TAG content, through transient co-expression of four avocado (Persea americana) genes, namely WRINKLED1 (WRI1), WRI2, diacylglycerol acyltransferases 1 (DGAT1), and phospholipid: diacylglycerol acyltransferase 1 (PDAT1) in Nicotiana benthamiana leaves. We hypothesize that stable expression of these “push–pull” avocado module genes in moss will increase storage lipid synthesis and redirect carbon flux toward TAG in vegetative biomass. Avocado cDNAs encoding the transcription factors PaWRI1 and PaWRI2 were cloned into Gateway-compatible expression vectors and introduced into moss protoplasts via PEG-mediated transformation. Stable transgenic moss lines expressing PaWRI1 and PaWRI2 were successfully generated. Total lipids from wild-type gametophores were extracted and analyzed by thin-layer chromatography (TLC) and gas chromatography–mass spectrometry (GC–MS) to establish a baseline lipid profile. Ongoing work is focused on evaluating TAG accumulation in the transgenic lines, as well as visualizing and quantifying lipid droplet formation using Nile Red staining and confocal fluorescence microscopy. In parallel, the avocado acyltransferase genes PaDGAT1 and PaPDAT1 are being cloned into Gateway-compatible expression vectors for subsequent introduction into moss, enabling future stacking of the full four-gene avocado lipid-biosynthesis module to further enhance TAG production. Collectively, these approaches will establish moss as a controllable, vegetative platform for plant oil engineering.
